In the course of screening for hyaluronidase (HAase) inhibitory agents, a new gamma-pyrone derivative, lepidepyrone, C8H10O5, was isolated from the cultured mycelium of the mushroom Neolentinus lepideus TMC-1102 as a major HAase inhibitory compound (IC50 3.3 mM). The structure of lepidepyrone was established on the basis of spectroscopic investigation.